The in vitro and in vivo Neuroprotective and Osteoprotective Effects of an Innovative Chinese Medicine Formula against Dementia and Osteoporosis

摘要

Health burden raising from ageing is becoming apparent in the last few decades. Currently, ageing-related degenerative diseases such as dementia (Alzheimer's disease [AD] and vascular dementia [VaD]) and osteoporosis are incurable that conventional medications can only slow down the progression of diseases. Traditional Chinese Medicines (TCM) such as Gastrodiae Rhizoma (Tianma) and Salviae Miltiorrhizae Radix Et Rhizoma (Danshen) have been reported to have neuroprotective and provide vascular tonifying effects, while Ligustri Lucidi Fructus (Nuzhenzi) reported to be osteoprotective. We hypothesized that this innovative formula could provide neuroprotection towards AD and VaD, and promote bone health. The objective of the present investigation was to study the effects of combination of three selected TCM herb aqueous extracts against AD and VaD, meanwhile tackling osteoporosis.;All individual TCM extracts demonstrated neuroprotective and osteoprotective effects with significant increased cell viability in in vitro Abeta-induced toxicity and H2O2-indcued oxidative stress assays. The optimal ratio of Danshen and Tianma (concentration ratio of 2:1) was determined with cell survival assays against Aâ- induced toxicity. Nuzhenzi was removed from the formula due to its relatively high cytotoxicity in cell culture. Thus, a novel herbal formula Danshen Tianma (DT) formula in the ratio of 2:1 was established. DT also constituted neuroprotective effects by inhibiting reactive oxygen species (ROS) activities, Aâ-induced apoptosis and acetylcholinesterase (AChE) activity. Osteoprotection of DT formula was revealed by enhancement in matrix mineralization and inhibition of osteoclasts differentiation.;The middle cerebral artery occlusion (MCAO) model indicated DT was able to decrease the neurological deficit and cerebral ischemic infarct volume. Further mechanistic study demonstrated DT could upregulate of anti-oxidative enzymes, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities; as well as downregulate of pro-inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha).;In the 5xFAD transgenic AD mice model, DT was found to improve the learning ability and spatial memory of the mice. These improvements were attributed to a diminished Abeta burden by inhibition of BACE1 activity, the suppression of glial activation and reduced neuroinflammation. On the other hand, DT formula also comprised osteoprotective effects with the efficacy of improvement in bone microarchitecture.;Our group has have established a novel passive mouse smoking-induced small vessel disease (SVD)-osteopenic model with attenuation of cerebral vessel density and femur bone microstructure. After DT treatment, smoking-induced cerebral vessel density reduction was alleviated and bone micro-architecture was improved via suppression of release of TNF-alpha and IL-6.;In conclusion, the current comprehensive study revealed the protection of DT against ageing-related degenerative disease, dementia and osteoporosis, via antioxidation, anti-inflammation, anti-apoptosis and anti-amyloidogenesis.